1. Field of the Invention
The invention relates to a Bioprinting station, to an assembly comprising such Bioprinting station and to a Bioprinting method.
In particular, the invention relates to Bioprinting (biological printing) station comprising:
a Bioprinting device adapted to deposit a pattern of biological material (including cells, cell aggregates, biomaterials, nanoparticles, drugs and other molecules having a biological effect on cells of a tissue, and others), onto an area of interest of a substrate, said area of interest having a feature (recess, specific morphology, optical feature, marking or other) that distinguishes said area of interest from a remaining part of the substrate, said Bioprinting device comprising:                at least one biological material dispenser adapted to dispense the biological material to be deposited, and        a positioning system adapted to receive the substrate and to position the area of interest with respect to the dispenser,        
an electronic control unit adapted to drive the dispenser and the positioning system relative to one another according to the pattern to be deposited.
Although not limited thereto, the invention has particularly advantageous applications in the treatment of loss of tissue architecture (including multiple cell types and matrix components precisely organized in three dimensions) caused for example by a trauma or a disease and which leads to loss of tissue function.
2. Description of Related Art
It has been found that such loss of tissue architecture could be treated through generation of biological tissues involving the use of engineering and material methods to obtain the appropriate combination of cells and the suitable biochemical and physicochemical factors which mimic both the micro-environment of cells and the micro-architecture of tissue in the body. In this context, tissue engineering which aims to provide for biological substitutes which restore, maintain or improve tissue function or a whole organ has been developed.
At first, living cells have been seeded onto biocompatible, and eventually biodegradable, scaffold and cultured in a bioreactor to lead to an initial cell population expanding into a tissue. With an appropriate scaffold which mimics the biological extracellular matrix, the developing tissue can adopt both the form and the function of the desired organ, and can be implanted into the body of patient.
In parallel with the aforementioned method, the building of three-dimensional (3D) biological structures by the technology of Bioprinting has been considered (“Application of laser printing to mammalian cells”, J. A. Barron, B. R. Ringeisen, H. Kim, B. J. Spargo, et D. B. Chrisey, Thin Solid Films, vol. 453-454, April. 2004, 383-387; “Quantification of the activity of biomolecules in microarrays obtained by direct laser transfer”, V. Dinca, A. Ranella, M. Farsari, D. Kafetzopoulos, M. Dinescu, A. Popescu, et C. Fotakis, Biomedical Microdevices, vol. 10, October. 2008, 719-25). Bioprinting consists in an automated, computer-aided layer-by-layer deposition, transfer and patterning of biological materials including cells and cell aggregates (“Organ printing: computer-aided jet-based 3D tissue engineering”, V. Mironov, T. Boland, T. Trusk, G. Forgacs, and R. R. Markwald, Trends in Biotechnology, vol. 21, April. 2003, 157-161; “Biofabrication: a 21st century manufacturing paradigm”, V. Mironov, T. Trusk, V. Kasyanov, S. Little, R. Swaja, et R. Markwald, Biofabrication, vol. 1, 2009, p. 022001; “Jet-based methods to print living cells”, B. R. Ringeisen, C. M. Othon, J. A. Barron, D. Young, et B. J. Spargo, Biotechnology Journal, vol. 1, September. 2006, 930-48). Recently, the definition of bioprinting was enlarged to “the use of computer-aided transfer processes for patterning and assembling living and non-living materials with a prescribed 2D or 3D organization in order to produce bio-engineered structures serving in regenerative medicine, pharmacokinetic and basic cell biology studies” (F. Guillemot, V. Mironov, M. Nakamura, Biofabrication, vol. 2, 2010).
To this end, commercially available inkjet printers have been redesigned (“Application of inkjet printing to tissue engineering”, T. Boland, T. Xu, B. Damon, and X. Cui, Biotechnology Journal, vol. 1, 2006, 910-917) or new ones built (“Biocompatible inkjet printing technique for designed seeding of individual living cells”, Makoto Nakamura, Akiko Kobayashi, Fumio Takagi, Akihiko Watanabe, Yuko Hiruma, Katsuhiro Ohuchi, Yasuhiko Iwasaki, Mikio Horie, Ikuo Morita, Setsuo Takatani, Tissue Eng 2006; “Delivery of human fibroblast cells by piezoelectric drop-on-demand inkjet printing”, Saunders R E, Gough J E, Derby B., Biomaterials 2008; 29: 193-203.) to pattern biological assemblies according to a computer-aided design template. Pressure-operated mechanical extruders such as bioplotters have also been developed to handle live cells and cell aggregates (“Tissue Engineering by Self-Assembly of Cells Printed into Topologically Defined Structures”, K. Jakab, C. Norotte, B. Damon, F. Marga, A. Neagu, C. L. Besch-Williford, A. Kachurin, K. H. Church, H. Park, V. Mironov, R. Markwald, G. Vunjak-Novakovic, and G. Forgacs, Tissue Engineering Part A, vol. 14, 2008, 413-421).
Parallel to these Bioprinting methods, laser-assisted printing has emerged as an alternative method for the assembly and micro-patterning of biological materials.
Laser-guided direct writing (LGDW) is a technique capable of trapping multiple cells in a laser beam and depositing them as a steady stream onto arbitrary non-absorbing surfaces (“Laser-guided direct writing for three-dimensional tissue engineering” Nahmias Y, Schwartz R E, Verfaillie C M, Odde D J, Biotechnol Bioeng 2005; 92: 129-36; “Micropatterning of living cells by laser-guided direct writing: application to fabrication of hepatic-endothelial sinusoid-like structures”, Yaakov Nahmias, David J. Odde, Nat Protoc 2006).
Laser-Assisted Bioprinting (LAB) is based on the laser-induced forward-transfer (LIFT) technique in which a pulsed laser is used to induce the transfer of biological material from a ribbon as a reservoir, formed of a layer of biological material spread onto an optically transparent quartz support, to a substrate in close proximity to or in contact with the ribbon (“Laser printing of pluripotent embryonal carcinoma cells”, Ringeisen B R, Kim H, Barron J A, Krizman D B, Chrisey D B, Jackman S, Auyeung R Y C, Spargo B J, Tissue Eng 2004; 10: 483-91).
Known Laser-Assisted Bioprinting methods comprise matrix assisted pulsed laser evaporation-direct write (MAPLE-DW) (“Application of laser printing to mammalian cells”, Barron J A, Ringeisen B R, Kim H, Spargo B J, Chrisey D B, Thin Solid Films 2004: 383-7), absorbing film assisted-LIFT (AFA-LIFT) (“Survival and proliferative ability of various living cell types after laser-induced forward transfer”, Bela Hopp, Tomi Smausz, Norbert Kresz, Norbert Barna, Zsolt Bor, Lajos Kolozsvari, Douglas B. Chrisey, Andras Szabo, Antal Nogradi, Tissue Eng 2006) and Laser-Assisted Bioprinting (LAB) (“Laser-Assisted Bioprinting: a novel technique for creating heterogeneous 3-dimensional cell patterns”, Barron J A, Wu P, Ladouceur H D, Ringeisen B R, Biomed Microdev 2004; 6: 139-47; “Laser printing of single cells: statistical analysis, cell viability, and stress”, Barron J A, Krizman D B, Ringeisen B R, Ann Biomed Eng 2005; 33: 121-30). Using LAB, under suitable irradiation conditions, and for liquids presenting a wide range of rheologies, the material can be deposited in the form of well-defined circular droplets with a high degree of spatial resolution.